LOCUS       sequence-417674-        6071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of the bright and photostable monomeric 
            StayGold fluorescent protein (E138D). Contains a N-terminal 10xHis 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6071)
  AUTHORS   Ivorra-Molla E, Akhuli D, McAndrew MBL, Scott W, Kumar L, Palani S, 
            Mishima M, Crow A, Balasubramanian MK
  TITLE     A monomeric StayGold fluorescent protein.
  JOURNAL   Nat Biotechnol. 2023 Dec 11. doi: 10.1038/s41587-023-02018-w.
  PUBMED    38081970
REFERENCE   2  (bases 1 to 6071)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2023 Dec 11. doi: 10.1038/s41587-023-02018-w."
COMMENT     Sequence Label: pcDNA3-10His-mStayGold (E138D)
FEATURES             Location/Qualifiers
     source          1..6071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(580..1371,1372..1440))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1203..1222
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1441..1545)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1620..1639)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1811..2190
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2191..2394
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2345..2365
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2439..2458
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2439..2457
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2471..2480
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2489..2518
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2525..3175
                     /codon_start=1
                     /product="bright and highly photostable monomeric green 
                     fluorescent protein (Ivorra-Molla et al., 2023)"
                     /label=mStayGold (E138D)
                     /note="contains the monomerizing E138D mutation"
                     /translation="MASTPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKGKYVCTSGKL
                     PMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGSIHAKHQHFM
                     KNGTYHNIVEFTGQDFKENSPVLTGDMNVSLPNDVQHIPRDDGVECPVTLLYPLLSDKS
                     KCVEAHQNTICKPLHNQPAPDVPYHWIRKQYTQSKDDTEERDHICQSETLEAHL"
     promoter        complement(3200..3218)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3201..3218)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3238..3255)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3244..3468
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3514..3942
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3601..3620)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3811..3832
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3951..3971)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3956..4285
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4136..4271
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4198..4217
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4352..5146
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4406..4425)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5016..5035
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5320..5441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5357..5376)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5411..5430
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5490..5506)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5490..5506)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5503..5525)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5514..5530
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5538..5544,5545..5562,5563..5568))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5583..5604
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5721..5738)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5892..6071,1..409))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5972..5991)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
       61 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      121 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      181 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      241 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      301 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      361 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      421 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      481 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      541 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
      601 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      661 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
      721 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
      781 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
      841 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
      901 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
      961 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1021 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1081 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1141 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1201 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1261 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1321 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1381 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1441 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     1501 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     1561 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     1621 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     1681 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     1741 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     1801 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     1861 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     1921 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     1981 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     2041 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     2101 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     2161 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2221 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2281 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2341 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
     2401 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
     2461 acccaagctt gccaccatgg gcagcagcca tcaccatcac catcaccatc accatcacgg
     2521 ttctatggcc agcacgccgt ttaagttcca gttaaagggg acaattaacg ggaaaagctt
     2581 tacagtcgag ggtgaagggg aagggaactc gcatgaaggt agtcataaag gtaagtatgt
     2641 gtgtacgagc ggaaagttgc cgatgtcttg ggcggctctt ggtacttcgt ttgggtacgg
     2701 aatgaaatac tatacaaaat acccgtcggg cctgaaaaat tggttccatg aggtgatgcc
     2761 cgaggggttc acgtatgatc gccacatcca gtacaaaggt gatggctcaa tccacgcaaa
     2821 gcaccaacac ttcatgaaga atgggactta tcataacatt gtcgagttta ctggccagga
     2881 cttcaaggaa aacagccccg tattgacagg tgacatgaat gtgagcttgc cgaacgacgt
     2941 gcagcatatt ccacgtgacg acggagttga atgtcccgtc acattgttat accccttgct
     3001 tagcgataaa tcaaagtgtg tcgaagcaca tcaaaacact atctgcaaac ctcttcataa
     3061 ccaacctgcg ccagatgtac cgtaccattg gattcgcaaa caatatacac agtctaagga
     3121 tgatacggag gaacgcgatc atatctgtca gtccgagacg ttagaagctc atctttaaca
     3181 tgcatctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct
     3241 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     3301 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     3361 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     3421 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     3481 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     3541 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3601 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3661 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3721 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3781 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3841 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3901 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     3961 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4021 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4081 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4141 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4201 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     4261 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     4321 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4381 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4441 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4501 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     4561 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4621 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4681 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4741 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4801 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4861 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4921 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4981 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     5041 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5101 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     5161 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     5221 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5281 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     5341 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5401 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     5461 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5521 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5581 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5641 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5701 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5761 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5821 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5881 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5941 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6001 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6061 ccttcgggaa g
//
